Refolding variant of bone morphogenetic protein-8

DNA molecules encoding mutant forms of bone morphogenetic proteins (BMP) are disclosed. The mutant forms of BMP can be produced bacterially and refolded to produce biologically active homodimers or heterodimers of BMP. A method of making such mutant BMPs is also disclosed.

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Claims

1. A recombinant DNA molecule comprising a nucleotide sequence encoding Bone Morphogenetic Protein-8 (BMP-8) as shown in SEQ ID NO:11, except that the nucleotide triplet at 187 to 189 has beenreplaced with a triplet encoding histidine.

2. A purified composition of Bone Morphogenetic Protein-8 (BMP-8) comprising the amino acid sequence of SEQ ID NO:12, wherein the serine at amino acid residue 63 has been replaced with histidine.

3. A purified composition of Bone Morphogenetic Protein (BMP) heterodimer comprising at least one mutant BMP encoded by a DNA molecule according to claim 1.

4. A purified composition of Bone Morphogenetic Protein (BMP) heterodimer comprising at least one mutant BMP according to claim 2.

Referenced Cited
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5013649 May 7, 1991 Wang et al.
5106748 April 21, 1992 Wozney et al.
5141905 August 25, 1992 Rosen et al.
5187076 February 16, 1993 Wozney et al.
Foreign Patent Documents
114 506 February 1989 EPX
433 225 September 1991 EPX
WO 91/18098 August 1991 WOX
9215323 September 1992 WOX
WO 93/00432 February 1993 WOX
Other references
  • King, "Genetic Analysis of Protein Folding Pathways" Bio/Technology 4:297-303 (Apr. 1986). Ozkaynak et al, "Osteogenic Protein-2" J. Biol. Chem. 267(35):25220-25227 (Dec. 1992). Kohno, Meth. Enzym. 185:187-195 (1990). Taniguchi et al., PNAS USA 77:5230-5233 (1980). Thies et al., J. Bone and Mineral Res. 5(2):305 (1990). Thies et al., Endocrinology 130:1318-1324 (1992). Sampath and Reddi, PNAS USA 80:6591-6595 (1983). Norrander et al., Gene 26:101-106 (1983). Sanger et al., J. Mol. Biol. 162:729-773 (1982). Takagi et al., Nucl. Acids. Res. 13:2063-2074 (1985). Dagert and Ehrlich, Gene 6:23 (1979). Tandon and Horowitz, J. Biol. Chem., 262:4486-4491 (1987).
Patent History
Patent number: 5756308
Type: Grant
Filed: Dec 21, 1994
Date of Patent: May 26, 1998
Assignee: Genetics Institute, Inc. (Cambridge, MA)
Inventors: Neil M. Wolfman (Dover, MA), John McCoy (Reading, MA)
Primary Examiner: Stephen G. Walsh
Assistant Examiner: Elizabeth C. Kemmerer
Attorneys: Steven R. Lazar, Thomas J. DesRosier
Application Number: 8/360,914
Classifications
Current U.S. Class: 435/691; 536/235; Synthesis Of Peptides (530/333); Hormones, E.g., Prolactin, Thymosin, Growth Factors, Etc. (530/399)
International Classification: C07K 1451; C12N 1512;